Qalabka CHO HCP ELISA

Bisadda No: HCP0032A

Xidhmada:96T

Habka difaaca jirka ee ELISA ee hal-tallaabo ah ayaa lagu isticmaalay baaritaankan.Tusaalooyinka ay ku jiraan CHOK1 HCP waxay isku mar la falgalaan HRP-ku calaamadsan orgiga ka hortagga CHOK1 antibody iyo anti-CHOK1 ee ku dahaarka saxanka ELISA.

Soo dir weydiin

Sharaxaada Alaabta

Taariikhda alaabta

Habka difaaca jirka ee ELISA ee hal-tallaabo ah ayaa lagu isticmaalay baaritaankan.Tusaalooyinka ay ku jiraan CHOK1 HCP waxay si isku mar ah ula falgalaan HRP-ku calaamadsan orgiga ka hortagga-CHOK1 iyo antibody-ka-hortagga CHOK1 ee dahaarka leh ee saxanka ELISA, ugu dambayntiina samaynaysa isku-dhafan sandwich ah oo ah antibody-ka-hortagga-HCP-ku calaamadsan.Antigen-antigen ee aan xidhnayn waxa laga saari karaa iyada oo la dhaqo saxanka ELISA.Substrate-ka TMB ayaa lagu daraa ceelka si loo helo falcelin ku filan.Horumarinta midabka waa la joojiyaa ka dib marka lagu daro xalka joogsiga, iyo OD ama qiimaha nuugista ee xalka falcelinta ee 450/650nm waxaa lagu akhriyaa akhristaha microplate.Qiimaha OD ama qiimaha nuugista ayaa u dhigma tusmada HCP ee xalka.Laga soo bilaabo tan, xoojinta HCP ee xalka waxaa lagu xisaabin karaa iyadoo loo eegayo qalooca caadiga ah.

Hore: Qalabka Genotyping Mouse HCR2021A

Xiga: RNA (dsRNA) labajibaaran ku xidhan ELISA KIT HCP0033A

Codsiga

Qalabkan waxa loo isticmaalaa in qiyaas ahaan lagu ogaado waxa ku jira CHOK1 hadhaaga borotiinka unugga martida loo yahay ee muunado.

Cxoogsheegayaal

S/N	Qayb	Xoog-saarid	Xaaladaha Kaydinta
1	Heerka CHOK1 HCP	0.5mg/ml	≤-20℃
2	Anti-CHO HCP-HRP	0.5mg/ml	≤–20℃, ka ilaali iftiinka
3	TMB	NA	2-8℃, ka ilaali iftiinka
4	20 × PBST 0.05%	NA	2-8℃
5	Jooji xal	NA	RT
6	xidhayaasha Microplate	NA	RT
7	BSA	NA	2-8℃
8	Taarikada dahaarka hore ee adsorption sare	NA	2-8℃

Qalabka Loo Baahan Yahay

Alaabta/qalabka	Wax-soo-saarka	Buug-gacmeedka
Akhristaha Microplate	Qalabka molecular	Spectra Max M5, M5e, ama wax u dhigma
Thermomixer	Eppendorf	Eppendorf/5355, ama wax u dhigma
Isku-darka Vortex	IKA	MS3 Digital, ama u dhigma

Kaydinta iyo xasilloonida

1.Gaadiidka at -25 ~ -15°C.

2.Xaaladaha kaydinta ayaa ah sida ku cad Shaxda 1;qaybaha 1-2 waxa lagu kaydiyaa ≤-20°C,5-6 waxa lagu kaydiyaa RT,3,4,7,8 waxa lagu kaydiyaa 2-8℃;muddada ansaxnimadu waa 12 bilood.

Xuduudaha alaabta

1.Dareenka: 1ng/ml

2.Qiyaasta ogaanshaha:3-100ng/ml

3.Saxnimada: Intra-ssay CV≤ 10%, CV≤ 15%

4.Caymiska HCP:>80%

5.Gaar ahaan: Xirmadani waa mid caalami ah maadaama ay si gaar ah ula falgasho CHOK1 HCP oo ka madax bannaan habka nadiifinta.

Diyaarinta reagent

1.PBST 0.05%

Qaado 15 ml 20×PBST 0.05%, lagu qasi jiray ddH₂O, oo laga sameeyay ilaa 300 ml.

2.1.0% BSA

Qaado 1g oo BSA ah dhalada oo ku qas 100 ml oo PBST 0.05% ah, si fiican isku walaaq ilaa ay si buuxda u milmaan, kuna kaydi 2-8°C.Badbaadiyaha la diyaariyey wuxuu shaqaynayaa 7 maalmood.Waxaa lagu talinayaa in la diyaariyo marka loo baahdo.

3.Xalka ogaanshaha 2μg/ml

Qaado 48μL oo ah 0.5 mg/mL Anti-CHO HCP-HRP oo ku milo 11,952μL ee 1% BSA si aad u hesho fiirsashada kama dambaysta ah ee 2μg/mL xal ogaanshaha.

4.QC iyo Diyaarinta Heerarka CHOK1 HCP

Tube Maya	Asalka dolution	Xoog-saarid ng/ml	Mugga μL	1% BSA Mugga μL	wadarta Mugga μL	Ugu dambayn xooga saarid ng/ml
A	Heerka	0.5mg/ml	10	490	500	10,000
B	A	10,000	50	450	500	1,000
S1	B	1.000	50	450	500	100
S2	S1	100	300	100	400	75
S3	S2	75	200	175	375	40
S4	S3	40	150	350	500	12
S5	S4	12	200	200	400	6
S6	S5	6	200	200	400	3
NC	NA	NA	NA	200	200	0
QC	S1	100	50	200	250	20

Shaxda: Diyaarinta QC iyo Heerarka

Habka Qiimaynta

1.U diyaari reagents sida ku cad "Diyaarinta Reagent" ee kore.

2.Qaado 50μL ee heerarka, muunado iyo QCs (tixraac shaxda 3) ceel kasta, ka dibna ku dar 100μL ee xalinta ogaanshaha (2μg / mL);Saxanka ku dabool daboolka, oo dhig saxanka ELISA heerkulbeegga.Ku dhaji 500rpm, 25± 3℃ 2 saacadood.

3.U rogo microplate-ka saxanka ku jira oo tuur xalka daahan.Pipette 300μL ee PBST 0.05% geli ceel kasta si aad u dhaqdo saxanka ELISA oo aad u tuurto xalka, oo ku celi dhaqista 3 jeer.U rog saxanka ku dul shukumaan warqad nadiif ah oo qalaji.

4.Ku dar 100μL substrate TMB ah (eeg Shaxda 1) ceel kasta, xidh saxanka ELISA, oo ku dhex daadi gudcurka 25± 3℃ 15 daqiiqo.

5.Pipette 100μL oo ah xalka joogsiga ceel kasta.

6.Ku cabbir nuugista mawjada dhererka 450/650nm oo leh akhristaha microplate.

7.Ku taxliilin xogta SoftMax ama software u dhigma.Qabo qalooca caadiga ah adiga oo isticmaalaya qaabka dib u noqoshada saadka oo afar-beeg ah.

Tusaalaha Curve Standard

FIIRO GAAR AH: Haddii xoogga HCP ee muunadku uu dhaafo xadka sare ee qalooca caadiga ah, waxa ay u baahan tahay in si habboon loogu qaso badhxiyaha ka hor inta aan la tijaabin.

XUSUUS

Xalka joogsiga waa 2M sulfuric acid, fadlan si taxadar leh u qabooji si aad uga fogaato daadinta!